Tsutsumi Victor

Publications (2)0 Total impact

• Article: Ultrastructural Evidences of Hepatitis B Infection in Human Liver Biopsies Disclose Complex Assembly and Morphogenesis Pathways for Hepatitis B Virus

  Falcon Viviana, Menéndez Ivon, Acosta-Rivero Nelson, Shibayama Mineko, Marıa-C de la Rosa, Jose Luna-Munoz, Miranda-Sanchez Magdalena, Jorge V Gavilondo, Lopez Deliana, Duenas-Carrera Santiago, [......], Vidal Eduardo, Arus-Soler Enrique, Silva Jose, Alvarez Felix, Emilio F. Acosta, Seoane Jesus, Morales-Grillo Juan, Penton Eduardo, Kouri Juan, Tsutsumi Victor
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  ABSTRACT: Despite of recent advances on acknowledge of hepatitis B virus (HBV) structure and biology, little is known about the morphogenesis and release of virions. In this study, the ultrastructural analysis of HBV components in liver biopsies from chronically HBV-infected patients disclosed complex assembly and morphogenesis pathways for HBV. HBV core (HBcAg) and surface (HBsAg) antigens were specifically identified in all liver biopsies from HVB-infected patients. HBcAgcontaining core-like particles 24-28 nm in diameter were observed both in nucleus and cytoplasm of hepatocytes. Dane’s-like particles were detected either budding to or into the lumen of ER close toHBsAg containing tubular structures. Besides, Dane’s-like particles were detected in different vesicular compartments resembling multivesicular endosomes. Other kind of enveloped HBV-like particles similar to the previously described cobra-shaped and horn-shaped particles were also observed in hepatocytes. Some of these particles were connected to the vesicle membrane through a stalk 20-22 nm in diameter. On the other hand, spherical subviral particles (SVP) were frequently observed in cytoplasmic vesicles. Moreover, a minor proportion of enveloped HBV-like particles budding through the plasma membrane to the extracellular space and bile canaliculi were detected.Interestingly, Dane’s-like particles in the bile canaliculi and entering into ductular-like cells were shown. Strikingly, Dane’s-like particles close to tubular structures and specific immunolabeling forHBcAg both in cytoplasm and nuclei of stellate-like cells were detected. Remarkably, HVB-like particles appeared entering hepatocytes from large cytoplasmic processes of fibrocytes raising theinteresting possibility of a cell to cell passage of HBV through direct transcellular migration.
  American Journal of Infectious Diseases. 01/2008;
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  Available from: scipub.org

  Article: Detection of HCV Components and Pathological Reactions in Apoptotic Hepatocytes from Chronically HCV-infected Patients

  Falcón Viviana, Acosta-Rivero Nelson, Shibayama Mineko, Luna-Munoz Jose, Miranda-Sanchez Magdalena, Gavilondo Jorge, María-C de la Rosa, Menéndez Ivón, Gra Bienvenido, García Waldo, Dueñas-Carrera Santiago, Silva Jose, Chinea Glay, Maritza González Bravo, Alvarez Felix, Morales Juan, Kouri Juan, Tsutsumi Victor
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  ABSTRACT: Analysis of Hepatitis C Virus (HCV)-infected hepatocytes at the cellular level may contribute to elucidate the mechanisms of HCV pathogenesis. In this work, the presence of HCV components and pathological reactions in apoptotic hepatocytes from chronic HCV-infected patients were studied by electron microscopy and confocal microscopy. Eight samples of liver biopsies from patients with chronic hepatitis C were studied by laser scanning confocal microscopy, Transmission Electron Microscopy (TEM) and Immunoelectron Microscopy (IEM). Data provide evidence for apoptosis of hepatocytes from HCV-infected liver biopsies during chronic HCV infection. Confirmation of this process was based on the morphological data by TEM including cell shrinkage; chromatin condensation; formation of apoptotic bodies; phagocytosis by neighbouring cells; and the presence of DNA fragmentation by TUNEL assay and caspase 3 activation. Interestingly, Hepatitis C core protein (HCcAg) was specifically immunolabeled in the rough endoplasmic reticulum, mitochondria as well as in the nucleus of apoptotic hepatocytes. In addition, E1 was specifically immunostained in the cytoplasm and in the mitochondria of some hepatocytes. The presence of Crystalloid Bodies (CB) similar to those observed in recombinant P. pastoris expressing HCcAg was observed in the cytoplasm of some hepatocytes. Immunogold labelling showed that HCcAg co-localized with these CB. In addition, structures forming a paracristalline array and particles with a diameter of 50 nm appeared in the mitochondria of some apoptotic hepatocytes. Moreover, unstructured large aggregates containing HCcAg similar to those detected at late stages of HCcAg expression in recombinant P. pastoris cells were frequently observed in damaged hepatocytes. Of note, these aggregates were specifically immunostained with anti-HCcAg. Data suggest the possibility for a direct role of these HCV-related structures as well as HCcAg and E1 in apoptosis and pathogenicity.
  American Journal of Infectious Diseases. 01/2005;